A rapid method for monitoring in vitro glycoengineering

PROJECT SUMMARY The glycosylation of therapeutic proteinssuch as antibodiesis well known to affect bioactivityhoweveruniform glycosylation is challenging to achieve when glycoproteins are produced in bioreactorsOne approach to obtaining well defined homogeneous glycosylation is to use glycosidases and glycosyl transferasesglycoenzymesto modify the glycosylation of the proteinpost productionThis approach is often referred to as in vitro glycoengineeringHoweverunlike traditional small molecule chemical synthesisthere are no convenient cost effective methods or kits for monitoring the progress of the enzyme reactions during glycoengineeringHerewe propose to develop an innovative technology called GlycoSense that will permit in vitro glycoenzyme reactions to be monitored in near real time using flow cytometry with bead based glycanspecific reagentsFlow cytometry was chosen as the preferred platform because the data are statistically robustthe equipment is relatively commonplacethe analyses consume very low quantities of sampleand it will enable GandD to leverage its proprietary experience in producing carbohydrate sensing reagentsIn this Phase I applicationwe will demonstrate the ability of our GlycoSense technology to monitor the effect of glycoenzymes applied to the glycoengineering of commercial therapeutic biologicsprovided by a major biopharmaceutical companyThese biologics were chosen for the present proof of concept studies because of their medical importance and because of the emerging potential of glycoengineering in the development of biosimilars and biobetters